The effect of HPV 16 E2 on gC1qR e pression, the p38 MAPK JNK signalling pathway and apoptosis in gC1qR silenced cervical squamous carcinoma cells To further e plore the effect of HPV 16 E2 on gC1qR e pression, the p38 MAPK JNK signalling pathway and apoptosis in gC1qR silenced cervical squamous carcin oma cells, C33a and SiHa Rapamycin WY-090217 cells were treated with unmodified media or HPV 16 E2 vector. After 72 h, the cells were transfected with 100 ng of gC1qR siRNA or 100 ng of negative siRNA. gC1qR gene and protein e pression levels were analysed by real time PCR and Western blot analysis. The results demonstrated that gC1qR mRNA and protein e pression levels were signifi cantly increased in the HPV 16 E2 vector group com pared with the unmodified media group.

However, there was no difference between the HPV 16 E2 gC1qR siRNA group and the unmodified media group. gC1qR e pression in the HPV 16 E2 gC1qR siRNA treated group was significantly lower than that in the HPV 16 E2 group. In contrast, gC1qR e pression was notably increased in the HPV 16 E2 negative siRNA group compared with the HPV 16 E2 gC1qR siRNA group. Phospho p38 MAPK and phospho JNK were assessed by Western blot analysis in cervical squamous carcin oma cells that were treated with unmodified media or HPV 16 E2 vector. After 72 h, the cells were transfected with 100 ng of gC1qR siRNA or 100 ng of negative siRNA. As shown in Figure 3C, phospho p38 MAPK and phospho JNK were significantly increased in the HPV 16 E2 group and the HPV 16 E2 negative siRNA group compared with the unmodified media group.

There was no difference in p p38 MAPK and p JNK protein e pression between the unmodified media group and the HPV 16 E2 gC1qR siRNA group. However, the phosphorylated p38 MAPK and JNK protein were notably lower in the HPV 16 E2 gC1qR siRNA group compared with the HPV 16 E2 negative siRNA group. C33a and SiHa cell apoptosis was assessed by flow cy tometry following treatment with unmodified media or HPV 16 E2 vector. After 72 h, GSK-3 the cells were transfected with 100 ng of gC1qR siRNA or 100 ng of negative siRNA. The cells were double stained with Anne in V and PI. Early and late apoptotic cells were distributed in the Q1 LR and Q1 UR regions, respect ively. Necrotic cells were located in the Q1 UL region.

Figure 3D shows that accumulated HPV 16 E2 increased the C33a and SiHa cell number in the Q1 LR and Q1 UR regions in the HPV 16 E2 vector group and the HPV 16 E2 negative siRNA group compared with the unmodified media group. However, the Q1 LR and Q1 UR regions in the HPV 16 E2 gC1qR siRNA vector transfected cells showed a notable decrease com pared with the HPV 16 E2 vector Cabozantinib transfected group. However, the apoptotic cells in the HPV 16 E2 gC1qR siRNA group were significantly decreased compared with the HPV 16 E2 negative siRNA group.